Purification and characterization of a novel fibrinolytic enzyme from the polychaete, Neanthes japonica (Iznka)

Deng, Zhihui; Wang, Shaohua; Qi, Li; Ji, Xue; Zhang, Lianzhi; Hong, Ming

doi:10.1016/j.biortech.2009.10.014

Cited by 48 publications

(21 citation statements)

References 25 publications

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“…6). This molecular mass is similar to those previously reported for fibrinolytic proteins from other annelids [18,40,41]. Finally, the purified D. sugokai serine protease was not cytotoxic to endothelial cells even when treated at high concentrations (Fig.…”

Section: Discussionsupporting

confidence: 89%

A Novel Thrombolytic and Anticoagulant Serine Protease from Polychaeta, Diopatra sugokai

Kim¹,

Shim²,

Yeon³

et al. 2018

Journal of Microbiology and Biotechnology

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Ischemic stroke can result from blockage of blood vessels, forming fibrin clots in the body and causing irreparable brain damage. Remedial thrombolytic agents or anticoagulants have been studied; however, because the FDA-approved tissue plasminogen activator has low efficacy and side effects, it is necessary to develop safer and more effective treatment candidates. This study aimed at assessing the fibrinolytic and anticoagulation features of a novel serine protease extracted and purified from , a polychaeta that inhabits tidal flats. The purified serine protease was obtained through ammonium sulfate precipitation, affinity chromatography, and ion-exchange chromatography. Its molecular size was identified via SDS-PAGE. To characterize its enzymatic activities, the protease activity at various pH and temperatures, and in the presence of various inhibitors, was measured via azocasein assay. Its fibrinolytic activity and anticoagulant effect were assessed by fibrin zymography, fibrin plate assay, and fibrinogenolytic activity assays. The novel 38 kDa serine protease had strong indirect thrombolytic activity rather than direct activity over broad pH (4-10) and temperature (37°C-70°C) ranges. In addition, the novel serine protease exhibited anticoagulant activity by degrading the α-, β-, and γ-chains of fibrinogen. In addition, it did not produce cytotoxicity in endothelial cells. Therefore, this newly isolated serine protease is worthy of further investigation as a novel alkaline serine protease for thrombolytic therapy against brain ischemia.

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Section: Discussionsupporting

confidence: 89%

A Novel Thrombolytic and Anticoagulant Serine Protease from Polychaeta, Diopatra sugokai

Kim¹,

Shim²,

Yeon³

et al. 2018

Journal of Microbiology and Biotechnology

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“…Therefore, there has been a continual search for safer and cost-effective fibrinolytic agent from diverse sources. To date, fibrinolytic agents have been isolated and characterised from different sources, including plants, animals, microbes and fermented foods (Deng et al 2010; Choi et al 2011). Microbes are considers as a lucrative source of fibrinolytic agents due to their diverse biochemical nature, feasibility of mass culture and ease in genetic manipulation.…”

Section: Introductionmentioning

confidence: 99%

Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species

Meshram

Saxena

2016

3 Biotech

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Cost-effective, fibrinolytic agents possessing least or no antigenic properties are much in demand for their prospective use as clot busters in thrombolytic therapy. The present communication explores the potential of 22 endophytic Botryosphaeria species isolated from stem of Aegle marmelos for their fibrinolytic potential. Only nine fungal isolates exhibited proteolytic activity out of which only four possessed plasminogen-independent fibrinolytic activity. The endophytic fungal isolate #1088 AMSTITYEL showed maximum in vitro proteolytic and fibrinolytic activity with a halo formation of 153.86 and 113.04 mm2, respectively. The partially purified protein exhibited a fibrinolytic activity equivalent to 6.51 U/ml of plasmin. SDS–PAGE of the dialysed fraction of #1088 AMSTITYEL resolved into six bands ranging from 26 to 80 kDa. Fibrin zymogram exhibited that a single band of molecular size ~80 KDa possessing the fibrinolytic activity. Furthermore, the bioactive isolate #1088 AMSTITYEL was identified as Lasiodiplodia pseudotheobromae using classical and molecular taxonomic tools.Electronic supplementary materialThe online version of this article (doi:10.1007/s13205-016-0428-4) contains supplementary material, which is available to authorized users.

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“…The size of the clear hydrolytic zone did not change in the presence of plasminogen, suggesting that protease was a plasmin-like protease, which could directly degrade fibrin but not plasminogen activators such as urokinase (UK), streptokinase (SK), and tissue plasminogen activator (tPA). Therefore, secondary effects such as platelet activation related to plasmin formation could be avoided [20]. This is a specific advantage of serralysin over clinically used plasminogen activators.…”

Section: Discussionmentioning

confidence: 99%

Statistical Optimization of Media Components for Production of Fibrinolytic Alkaline Metalloproteases from Xenorhabdus indica KB-3

Pranaw

Singh

Dutta

et al. 2014

Biotechnology Research International

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Xenorhabdus indica KB-3, a well-known protease producer, was isolated from its entomopathogenic nematode symbiont Steinernema thermophilum. Since medium constituents are critical to the protease production, the chemical components of the selected medium (soya casein digest broth) were optimized by rotatable central composite design (RCCD) using response surface methodology (RSM). The effects of all five chemical components (considered as independent variables), namely tryptone, soya peptone, dextrose, NaCl, and dipotassium phosphate, on protease production (dependent variable) were studied, and it was found that tryptone and dextrose had maximum influence on protease production. The protease production was increased significantly by 66.31% under optimal medium conditions (tryptone—5.71, soya peptone—4.9, dextrose—1.45, NaCl—6.08, and dipotassium phosphate—0.47 in g/L). To best of knowledge, there are no reports on optimization of medium component for protease production by X. indica KB-3 using RSM and their application in fibrinolysis. This study will be useful for industrial processes for production of protease enzyme from X. indica KB-3 for its application in the field of agriculture and medicine.

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Purification and characterization of a novel fibrinolytic enzyme from the polychaete, Neanthes japonica (Iznka)

Cited by 48 publications

References 25 publications

A Novel Thrombolytic and Anticoagulant Serine Protease from Polychaeta, Diopatra sugokai

A Novel Thrombolytic and Anticoagulant Serine Protease from Polychaeta, Diopatra sugokai

Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species

Statistical Optimization of Media Components for Production of Fibrinolytic Alkaline Metalloproteases from Xenorhabdus indica KB-3

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